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J. Biol. Chem., Vol. 279, Issue 37, 38125-38133, September 10, 2004

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Phospholipase D Isozymes Mediate Epigallocatechin Gallate-induced Cyclooxygenase-2 Expression in Astrocyte Cells^*

Shi Yeon Kim, Bong-Hyun Ahn, Kyoung-Jin Min¶, Young Han Lee||, Eun-hye Joe¶, and Do Sik Min**

From the Department of Physiology, College of Medicine, The Catholic University of Korea, Seoul 137-701, Korea, the ^¶Department of Pharmacology, Ajou University, Suwon 44-721, Korea, and the ^||Division of Molecular & Life Sciences, College of Science & Technology, Hanyang University, Ansan 426-791, Korea

Little is known about the effect of epigallocatechin-3 gallate (EGCG), a major constituent of green tea, on the expression of cyclooxygenase (COX)-2. Here, we studied the role of phospholipase D (PLD) isozymes in EGCG-induced COX-2 expression. Stimulation of human astrocytoma cells (U87) with EGCG induced formation of phosphatidylbutanol, a specific product of PLD activity, and synthesis of COX-2 protein and its product, prostaglandin E₂ (PGE₂). Pretreatment of cells with 1-butanol, but not 3-butanol, suppressed EGCG-induced COX-2 expression and PGE synthesis. Furthermore, evidence that PLD was involved in EGCG-induced COX-2 expression was provided by the observations that COX-2 expression was stimulated by overexpression of PLD1 or PLD2 isozymes and treatment with phosphatidic acid (PA), and that prevention of PA dephosphorylation by 1-propranolol significantly potentiated COX-2 expression induced by EGCG. EGCG induced activation of p38 mitogen-activated protein kinase (p38 MAPK), and specific inhibition of p38 MAPK dramatically abolished EGCG-induced PLD activation, COX-2 expression, and PGE₂ formation. Moreover, protein kinase C (PKC) inhibition suppressed EGCG-induced p38 MAPK activation, COX-2 expression, and PGE₂ accumulation. The same pathways as those obtained ²in the astrocytoma cells were active in primary rat astrocytes, suggesting the relevance of the findings. Collectively, our results demonstrate for the first time that PLD isozymes mediate EGCG-induced COX-2 expression through PKC and p38 in immortalized astroglial line and normal astrocyte cells.

Received for publication, February 25, 2004 , and in revised form, June 18, 2004.

^* This work was supported by the Korea Research Foundation Grant (KRF-2003-015-E00060). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Both authors contributed equally to this work.

^** Present address: Dept. of Molecular Biology, College of Natural Science, Pusan National University, Busan 609-735, Korea. To whom correspondence should be addressed. Tel.: 82-51-510-1775; Fax: 82-51-513-9258.

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